(1) Field of the Invention
The present invention relates to a method and apparatus for the controlled actuation of a microscope, in general, and to controlled actuation of a laser scanning microscope having multiple light sources, in particular.
(2) Description of Related Art
Confocal microscopy is, among other things, the tool for defined controlled actuation of micro-objects. Based on that, numerous methods for examination and influencing of microscopic objects were proposed, thus, for instance, by Denk in U.S. Pat. No. 5,034,613, by Liu in U.S. Pat. No. 6,159,749, or by Karl Otto Greulich in “Micromanipulation by Light in Biology and Medicine” in 1999.
A combination comprising an image-forming point scanning or line scanning system and a “manipulator” system is increasingly finding more and more interest in professional circles.
The interest in the observation and analysis of fast microscopic processes has brought forth new devices and methods (for example Carl Zeiss Line Scanner LSM 5 LIVE), which, in combination with the above mentioned methods of manipulation, lead to new insights. Thereby, the simultaneous microscopic observation of radiation-induced manipulation of samples with spatial resolution by means of a suitable imaging system stands especially in the foreground (See for example U.S. Pat. No. 6,094,300 and DE 102004034987 A1). Therefore modern microscopes attempt to offer as many flexible and optically equivalent decoupling and coupling ports as possible (See: DE 102004016433 A1).
The availability at the same time of at least two coupling ports for independent scan systems is thereby of special importance in order to avoid limitations in temporal resolution due to the slowness of mechanical switching processes. Besides the tube interface, other coupling ports on the sides of the microscope stand are possible (preferably in the extended infinite space between the microscope objective and the tube lens; the so-called “sideports”) as well as on the rear side of the stand (typically optically modified incident light axis or transmitted light axis with suitable tube lens; the “rearports”) as well as on the bottom side (the “baseport”).
Thereby, arrangements with a common direction of the incident light (either reflected or transmitted light) or with a direction opposite to the incident light (transmitted light and reflected light) are possible in principle. Apart from the viewpoint of the applicability, a common direction of incidence is frequently preferred from the device-technical viewpoint. In that case, use of at least one element is necessary, which combines the beam paths of both devices in the space between the scanners of the scan systems that are to be operated simultaneously and the objective. Thereby, according to the state-of-the-art, a diverse variety of beam-combining elements are conceivable, such as, for example, optomechanical components, like suitably coated beam combiner flat plates and beam combiner wedges, beam combiner cubes and polarizing splitters. Conceivable are further beam combining acousto-optical modulators and deflectors.